CN1481360A - Muscarinic antagonists - Google Patents

Abstract

Amide derivatives of 1,4 di-substituted piperidine compounds of the formula or a pharmaceutically acceptable salt, ester or solvate thereof, wherein R<1> is optionally substituted cycloalkyl, cycloalkylalkyl, aryl, arylalkyl or heteroaryl; R<2> is H, alkyl, or optionally substituted cycloalkyl, cycloalkylalkyl, heterocycloalkyl, bridged cycloalkyl, or bridged heterocycloalkyl; R<3> is alkyl or -CH2OH; and R4 is H or alkyl; are muscarinic antagonists useful for treating cognitive disorders such as Alzheimer's disease. Pharmaceutical compositions and methods of preparation are also disclosed.

Description

Muscarinic antagonists

Background of the invention

The present invention relates to an amide derivative of 1,4-di-substituted piperidine for treating cognitive impairment, a pharmaceutical composition containing the compound, a treatment method using the compound, and the combined use of the compound and an acetylcholinesterase inhibitor the use of.

Piperidine-derived muscarinic antagonists for use in the treatment of cognitive disorders such as Alzheimer's disease are disclosed in U.S. Patent No. 6,037,352, and in particular U.S. Patent No. 6,037,352 discloses a compound of the general formula wherein Y is CH; Z is N; X is -NHCO-; R is substituted benzyl; R ¹ and R ²¹ are each H; R ³ , R ⁴ , R ²⁷ and R ²⁸ are hydrogen; and R ² is Cycloalkyl. Similar compounds in which the benzene ring is substituted with a pyridyl ring are also disclosed in US Patent No. 6,066,636. The compounds of the present invention are selected inventions of US Patent Nos. 6,037,352 and 6,066,636.

Summary of the invention

其药学上可接受的盐、酯或其溶剂化物，其中The present invention relates to the compound of following structural formula I or

Its pharmaceutically acceptable salt, ester or solvate thereof, wherein

R ¹ is R ⁵ -(C ₃ -C ₈ )cycloalkyl, R ⁵ -(C ₃ -C ₈ )cycloalkyl(C ₁ -C ₆ )alkyl, R ⁵ -aryl, R ⁵ -aryl -(C ₁ -C ₆ )alkyl or R ⁵ -heteroaryl;

R ² is H, (C ₁ -C ₆ ) alkyl, R ⁶ -(C ₃ -C ₈ ) cycloalkyl, R ⁶ -(C ₃ -C ₈ )-cycloalkyl-(C ₁ -C ₆ )-alkyl, R ⁶ -heterocycloalkyl, R ⁶ -(C ₆ -C ₁₀ ) bridged cycloalkyl, or R ⁶ -bridged heterocycloalkyl;

R ³ is C ₁ -C ₆ alkyl or -CH ₂ OH;

R ⁴ is H or C ₁ -C ₆ alkyl;

R ⁵ is independently selected from the group consisting of H, C ₁ -C ₆ alkyl, halogen, -OH, C ₁ -C ₆ -alkoxy, -CF ₃ , -CN, -CO ₂ R ⁴ , -CONHR ⁴ , - 1-4 substituents in SO ₂ NHR ⁴ , -NHSO ₂ R ⁴ and -NHC(O)R ⁴ ; and

R ⁶ is independently selected from the group consisting of H, C ₁ -C ₆ alkyl, halogen, -OH, C ₁ -C ₆ -alkoxy, -CF ₃ , -NH ₂ , (C ₁ -C ₆ )alkylamino , phenyl, C ₁ -C ₂ alkylenedioxy, and 1-4 substituents of (C ₁ -C ₆ ) alkoxycarbonyl.

Another aspect of the present invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I in a pharmaceutically acceptable carrier. The present invention also relates to methods of treating cognitive disorders or neurological diseases using compounds of formula I and pharmaceutical compositions comprising compounds of formula I, comprising administering an effective amount of the compounds or compositions of the present invention to a mammal in need of such treatment.

Another aspect of the present invention relates to a method for treating cognitive disorders or neurological diseases, comprising administering an effective amount of a combination of a compound of formula I and an acetylcholinesterase inhibitor to a mammal according to the need for the treatment.

A final aspect of the present invention relates to a kit for the treatment of cognitive disorders or neurological diseases, comprising single packaged pharmaceutical compositions for use in combination in separate containers, one of which contains formula I in a pharmaceutically acceptable carrier The compound, the second container is an acetylcholinesterase inhibitor in a pharmaceutically acceptable carrier, and the combined amount thereof is an effective amount.

Detailed Description of the Invention

Referring to formula I above, a preferred group of compounds is wherein R ¹ is R ⁵ -phenyl or R ⁵ -cyclohexyl. R ⁵ is preferably H, halogen or C ₁ -C ₆ alkyl, more preferably H, F, or -CH ₃ .

Another group of preferred compounds is wherein R ² is R ⁶ -C ₃ -C ₈ cycloalkyl, especially R ⁶ -C ₅ -C ₇ cycloalkyl, R ⁶ is preferably H or C ₁ -C ₆ alkyl .

R ³ is preferably —CH ₃ , and R ⁴ is preferably H.

Compared to, inter alia, the compounds disclosed in US Pat. No. 6,037,352 or 6,066,636, neither of which contains an ^R3 moiety, the compounds of the present invention are more selective than m2.

The term "alkyl" as used herein represents a straight or branched saturated hydrocarbon chain having the specified number of carbon atoms. If the number of carbon atoms is not listed, for example using the term lower alkyl, it may be from 1 to 6 carbons.

"Cycloalkyl" represents a saturated carbocyclic ring having 3 to 8 carbon atoms. A bridged cycloalkyl group refers to a cycloalkyl group in which two non-adjacent ring members are linked by a C ₁ -C ₂ alkyl chain.

The term "heterocycloalkyl" refers to a 4- to 7-membered saturated ring comprising 1 to 3 heteroatoms independently selected from the group comprising -O-, -S-, and ^-NR7- , wherein ^R7 is H or C ₁ -C ₆ alkyl, and wherein the remaining ring members are carbon. When the heterocycle includes more than one heteroatom, no ring is formed when there are adjacent oxygen atoms, adjacent sulfur atoms, or three consecutive heteroatoms. Examples of heterocycles are tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, thiomorpholinyl, and piperazinyl. A bridged heterocycloalkyl refers to one in which two non-adjacent carbonyl ring members are linked by a C ₁ -C ₂ alkyl group.

Halogen represents fluorine, chlorine, bromine or iodine.

Aryl represents phenyl or naphthyl.

Heteroaryl means a 5- or 6-membered aromatic ring comprising 1 to 3 independently selected heteroatoms comprising -O-, -S- and -N=, with the proviso that the ring does not contain adjacent oxygen and/or or a sulfur atom. Examples of heteroaryl are pyridyl, isoxazolyl, oxadiazolyl, furyl, pyrrolyl, thienyl, imidazolyl, pyrazolyl, tetrazolyl, triazolyl, thiadiazolyl, pyrazine group, pyrimidinyl, pyridazinyl, and triazolyl. All positional isomers are included, such as 2-pyridyl, 3-pyridyl and 4-pyridyl.

When there is more than one variable in the formula, eg R ⁵ , then more than one variation can be independently selected from the definition of variation.

The compounds of the invention have at least one asymmetric carbon atom, ie the carbon atom to which ^R3 is attached. All isomers including diastereomers, enantiomers and rotamers are part of the present invention. The present invention encompasses the d and 1 isomers in pure form and as premixtures of racemic mixtures. Isomers may also be prepared using conventional techniques for the reaction of optically pure or optically enriched starting materials, or for the separation of isomers of compounds of formula I. The preferred stereochemistry of the compound of the present invention is shown in formula IA:

Compounds of formula I may exist in unsolvated or solvated forms, including hydrated forms. In general, solvated forms (including pharmaceutically acceptable solvents such as water, ethanol, etc.) are equivalent to unsolvated forms for the purposes of the present invention.

The compounds of formula I can form pharmaceutically acceptable salts with organic and inorganic acids. Examples of suitable salt-forming acids are hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, citric acid, malonic acid, salicylic acid, malic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, methanesulfonic acid, and Other inorganic acids and carboxylic acids known to those skilled in the art. The above salts are produced in the usual manner by contacting the free base form with a sufficient amount of the desired acid. The free base form can be regenerated by treating the salt with a suitable dilute aqueous alkaline solution, such as sodium hydroxide, potassium carbonate, ammonia or sodium bicarbonate in aqueous solution. The free base form and its respective salt form differ somewhat in certain physical properties such as solubility in polar solvents, but for the purposes of the present invention a salt is another equivalent of its respective free base form.

Those skilled in the art can use known methods to prepare the compound of formula I, such as using the steps disclosed in US6,037,352, which is used as a reference of the present invention, or using parallel synthesis or combinatorial chemistry methods. It will be appreciated by those skilled in the art that other methods may also be used and that these methods may be suitably modified to prepare other compounds within the scope of compounds of formula I.

The compound of formula I above was prepared using the solid phase synthetic procedure as shown in Reaction Scheme 1 below, wherein Me is methyl and FMOC is 9-fluorenylmethoxycarbonyl. Reaction scheme 1

The synthesis of Reaction Scheme 1 can be accomplished by reacting 9-BBN with an alkene such as 2, followed by Suzuki coupling with an aryl halide such as 1 to give compound 3. Hydrolysis of ester 3 followed by removal of N-Boc afforded amino acid intermediate 5, which was protected by treatment with FmocOSU. This product is then converted to acid chloride 6 by treatment with reagents such as POCl ₃ or oxalyl chloride.

Amines (R ¹ CHR ³ NH ₂ ) were bonded to resin-bound aldehydes such as Argopore-MB-CHO resin (Argonaut Corporation, San Carlos, CA) by reductive alkylation with sodium triacetoxyborohydride. Subsequent acylation of the resin-bound amine (resin 1) with an activated acid such as acid chloride 7 affords resin 2. Deprotection of the N-Fmoc group followed by reductive alkylation with aldehydes or ketones, or reaction with aldehydes followed by treatment with Grignard reagents, or reaction with the appropriate methanesulfonate or alkyl halide to give resin bound The intermediate is treated with TFA to obtain the compound of formula I.

Compounds of formula I can also be prepared by general methods of synthetic chemistry. For example, compounds of formula Ia wherein R ¹ is R ⁵ -phenyl, R ³ is -CH ₃ , and R ⁴ is hydrogen can be prepared as described in Reaction Scheme 2:

Reaction flow 2

Reaction of an amine such as 8 with an activated carboxylic acid such as an acid chloride 9 in the presence of a base such as pyridine or triethylamine affords amides of type 10. Treatment with an acid such as TFA or hydrochloric acid affords compound 11. Derivatization of the piperidine nitrogen of compound 11 by reductive alkylation using an aldehyde or ketone in the presence of a reducing agent such as sodium triacetoxyborohydride, or alternatively reaction with an aldehyde followed by treatment with a Grignard reagent affords compounds of type Ia. Yet another method involves the reaction of amine 11 with the appropriate methanesulfonate or alkyl halide in the presence of a base.

The starting materials of formulas 7, 8 and 9 are known in the art, or prepared by methods known in the art, such as using ketones and aldehydes via reductive alkylation with alkyl halides or tosylates or introducing R ² through alkylation.

After the above reaction, if necessary or desired, one or more of the following steps can be carried out: (a) removing any protecting group from the prepared compound; (b) converting the prepared compound into pharmaceutically acceptable salts, esters and and/or a solvate; (c) converting the prepared compound of formula I into another compound of formula I; and (d) isolating the compound of formula I, including isolating the stereoisomers of formula I.

According to the above reaction sequence, those skilled in the art will be able to select the raw materials required for the preparation of any compound of formula I.

Compounds of formula I exhibit selective m2 muscarinic antagonist activity which has been corrected for pharmacological activity to treat cognitive disorders and/or their symptoms. Examples of cognitive disorders are Alzheimer's disease and aging disease, which can improve memory and learning ability after treatment.

Compounds of formula I are pharmaceutically active in a test procedure to demonstrate m1 and m2 muscarinic antagonist activity, the test method being described below. muscarinic binding activity

Compounds of the invention were tested for their ability to inhibit binding to cloned human ml, m2, m3, m4, and m5 muscarinic receptor subtypes. The source of receptors in this study was the membrane of a stably transferred CHO cell line, which expresses each receptor subtype. After growth, cells were pelleted and then homogenized using a Polytron in 50 volumes of cold 10 mM Na/K phosphate buffer, pH 7.4 (buffer B). The homogenate was also centrifuged at 40,000 xg for 20 minutes at 4°C. The resulting suspension was discarded and the pellet was resuspended in buffer B to a final concentration of 20 mg wet tissue/ml. Films were stored at -80°C until used in the binding assay described below.

Binding to cloned human muscarinic receptors was performed using ³ H-quinuclidinyl benzidine (QNB) (Watson et al., 1986). Briefly, films (approximately 8, 20 and 14 micrograms of protein were analyzed for films containing m1, m2 and m4, respectively) were incubated with ³ H-QNB (final concentration 100-200 PM) and incubated at 25°C for 90 minutes Bring the unlabeled drug concentration to a final volume of 2 mL. Non-specific binding was analyzed in the presence of 1 microliter of atropine. Cultures were terminated in GF/B glass fiber filters by vacuum filtration using Skatron filter units, and the filters were washed with cold 10 mM Na/K phosphate buffer (pH 7.4). The Scintillation mix was added to the filter and the bottle was incubated overnight. Bound radioligand was quantified in a liquid scintillation counter (efficiency 50%). The resulting data were analyzed for _IC50 values (ie the concentration of compound required to inhibit binding by 50%) using the EBDA computer program (McPherson, 1985). Affinity values (K _i ) were then determined using the following formula (Cheng and Prusoff, 1973):

Thus, a lower _Ki indicates greater binding affinity.

To determine the degree of selectivity of a compound for binding to the m2 receptor, the _Ki value for the m1 receptor is divided by the _Ki value for the m2 receptor. Higher values indicate greater selectivity for binding to m2 muscarinic receptors. microdialysis method

Compounds were shown to function as m2 antagonists using the following method.

Surgery : For three studies, male Spraque-Dawley rats (250-350 g) were anesthetized with sodium pentobarbital (54 mg/kg ip.) and placed on a Kopf Slerotaxic apparatus . The skull was exposed and a hole was made through 0.2 mm anterior to the dura and 3.0 mm lateral to the bregma. With coordination, a guide cannula was placed at the epidural edge through the burr opening, lowered vertically to a depth of 2.5 mm, and permanently fixed with dental plaster to the bone screws. After surgery, rats were dosed with ampicillin (40 mg/kg ip) and housed individually in modified cages. Allow about 3 to 7 days to recover before performing the microdialysis procedure.

Microdialysis : All equipment and instruments used for in vivo microdialysis were obtained from the company Bioanalytical Systems (BAS). The microdialysis procedure consisted of inserting a needle-like irritable probe (CMA/12.3 mm x 0.5 mm) through a thin guide cannula to a depth of 3 mm beyond the cannula-tip stripe. The probe is pre-tubed to a microsyringe pump (CMA/100). Rats were captured, tethered and then probed, and placed in large clear plastic glass basins with grass nests and access to food and water. The probe has been treated with Ringer's buffer (NaCl 147mM; KCl 3.0mM; CaCl ₂ 1.2mM; MgCl ₂ 1.0mM) perfusion 2μl/min. To achieve a stable baseline reading, the microdialysis was run for 90 minutes before collecting the fractions. Fractions (20 microliters) were obtained at 10 minute intervals over a 3 hour period using a cryocollector (CMA/170 or 200). Four to five baseline fractions are collected prior to dosing the animals with the test drug or combination. After collection, each rat was necropsied to determine the accuracy of probe placement.

Acetylcholine (ACh) analysis : HPLC/electrochemical detection/measurement of ACh concentration in samples collected by microdialysis. Samples were automatically injected (Waters 712 frozen sample processor) onto a polymer analysis HPLC column (BAS, MF-6150) and eluted with 50 mM Na ₂ HPO ₄ (pH 8.5). To avoid bacterial growth, Kathon CG reagent (0.005%) (BAS) was included in the mobile phase. The eluate from the analytical column, containing the separated ACh and choline, immediately passed through the Immobilized Enzyme Reactor Cassette (BAS, MF-6151 ) connected to the outlet of the column. The reactor contains acetylcholinesterase and choline oxidase, which are covalently bonded to the polymer backbone. The reaction of the above enzymes to ACh and choline resulted in a stoichiometric yield of hydrogen peroxide, which was detected electrochemically using a Waters 460 detector equipped with a platinum electrode and operated at a potential of 500 milliwatts. Data were acquired using an IBM Model 70 computer equipped with a microchannel IEEE board. Peak integration and quantification were accomplished using "Maxima" chromatography software (Waters Corporation). The total retention time for each sample was 11 minutes at a flow rate of 1 mL/min. The retention times of acetylcholine and choline were 6.5 and 7.8 minutes, respectively. To track and correct for possible changes in detector sensitivity during chromatography, ACh standards were added at the beginning, middle and end of each sample test.

The increase in ACh is consistent with a presynaptic m2 receptor antagonism mechanism. microsomal stability

A solution of the compound of formula I with a final substrate concentration of 0.5 mg/ml, and a solution of human, macaque or rat liver microsomes with a final P450 concentration of 0.18, 0.175 and 0.25 nmol/ml in 0.1 M potassium phosphate buffer at pH 7.4 Incubate in a 96-well micro-petri dish at 37°C for 3 minutes in a shaking water bath. A cofactor solution containing MgCl ₂ , glucose-6-phosphate, NADPH and glucose-6-phosphate dehydrogenase (half of the total incubation volume/sample) was added to each sample, and the entire incubation mixture was incubated for 0 and 30 minutes (n=3 samples incubated for each compound). After each time point, an equal volume of _CH3CN was added. Samples were mixed by shaking and the dishes were centrifuged at 3000 rpm for 20 minutes. The suspension is analyzed for the parent compound and/or metabolites by liquid chromatography mass spectrometry (LCMS) using appropriate analytical methods.

For the compounds of the present invention, the following ranges of muscarinic antagonist activity were found:

m1: 20-2000nM, the preferred compound is before 200-1000nM.

m2: 1-500nM, preferred compounds are <50nM, more preferably <10nM.

According to microsomal stability analysis, the results of the compound of Example 2 are as follows (residual % after 30 minutes): rat-79%, monkey-80%, human-80%.

In the aspect of the present invention in combination with a compound of formula I and an acetylcholinesterase inhibitor, examples of acetylcholinesterase inhibitors are donepezil, heptylphysostigmine, tacrine (tacrine), rivastigmine and galantamine.

For preparing pharmaceutical compositions from the compounds described by this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories. Powders and tablets may contain from about 5 to about 95% active ingredient. Suitable solid carriers are well known in the art, for example magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods for making various compositions can be found in Easton, PA, A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co.

Liquid form preparations include solutions, suspensions and emulsions. Examples are water or water-propylene glycol solutions for parenteral injection, or for oral solutions, suspensions or emulsions sweeteners or opacifying agents may be added. Liquid preparations also include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas such as nitrogen.

Also included are solid preparations, which are intended to be converted, shortly before use, to liquids for oral or parenteral administration. Such liquids include solutions, suspensions and emulsions.

The compounds of the invention may also be delivered transdermally. Transdermal compositions may be creams, paints, aerosols and/or emulsions, and may include transdermal patches on a base or carrier, as is generally common in the art.

Preferably the compounds are administered orally.

Preferred pharmaceutical formulations are in unit dosage form. In such form, the preparation is subdivided into suitably sized unit doses containing appropriate quantities of the active ingredient (eg, an effective amount) to achieve the desired purpose.

The amount of active ingredient in a unit dose of the preparation can be varied or adjusted from about 1 mg to about 100 mg, preferably from about 1 mg to about 50 mg, more preferably from about 1 mg to about 25 mg, depending on the specific application.

The actual dosage employed will vary according to the requirements of the patient and the severity of the condition being treated. The appropriate dosage for a particular situation will be determined by those skilled in the art. For convenience, the total daily dosage may be divided and administered in portions throughout the day as required.

The dosage and frequency of administration of the compound of the present invention and/or its pharmaceutically acceptable salt will be adjusted according to the clinical conditions such as the patient's age, symptoms and body size and the severity of the symptoms to be treated. The generally recommended daily dosage for oral administration is about 1 mg/day to about 300 mg/day, preferably 1 mg/day to about 50 mg/day, and is divided into two or four doses.

When the compound of formula I is used in combination with an acetylcholinesterase inhibitor to treat cognitive disorders, the two active ingredients can be administered simultaneously or sequentially, or the compound of formula I and acetylcholinesterase can be contained in a pharmaceutically acceptable carrier. Administration of a single pharmaceutical composition of a cholinesterase inhibitor. The components of the combination can be administered individually or together in any common oral or parenteral preparation form, such as capsules, tablets, powders, cachets, suspensions, solutions, suppositories, intranasal sprays, etc. The dose of acetylcholinesterase inhibitors may range from 0.001 to 100 mg/kg body weight.

The invention disclosed herein is illustrated by the following devices and examples, which should not limit the scope of the invention. Routes of other mechanisms and similar structures will be apparent to those skilled in the art. The following terms are abbreviated: room temperature (rt); 9-bromobicyclo[3.3.1]nonane (9-BBN); ethyl acetate (EtOAc); tetrafluoroacetic acid (TFA); tetrahydrofuran (THF); N-(9-fluorenylmethoxycarbonyl)hydroxysuccinimide (FMOC-OSuc); 1-(3-dimethylaminopropyl)-3-ethylcarbodiene Amine monohydrochloride (EDCI); 4-dimethylaminopyridine (DMAP); diethylazodicarboxylate (DEAD); and dichloroethane (EDC).

见上述的反应流程1Preparation Example 1

See reaction scheme 1 above

Starting material (2) (1 g) was mixed with 9-BBN (10.2 mL of a 0.5M solution in THF), left under nitrogen and heated to reflux for 1 h. Methyl 4-bromobenzoate (1.09 g), potassium carbonate (0.84 g), _PdCl2 (dppf) (0.21 g), _Ph3As (0.155 g), DMF (7 mL) and water (1.1 mL) were added In the cooled above solution, and heated at 65°C for 3 hours. The reaction mixture was poured into ice water, extracted with ethyl acetate, and the organic layer was purified by flash chromatography (hexane:ethyl acetate (90:10)) to obtain compound (3) (1.1 g). Compound (3) (1.1 g) was dissolved in methanol (20 mL), and lithium hydroxide (0.2 g) and water (7.5 mL) were added. After heating to reflux for 1 h, the reaction mixture was cooled, methanol was removed in vacuo, and the mixture was acidified with hydrochloric acid. The solid was collected by filtration and dried under vacuum to obtain compound (4). (4) was dissolved in 4M hydrochloric acid in dioxane (35 mL) and stirred for 1.5 hours. Diethyl ether was added and compound (5) (0.67 g) was collected by filtration. Compound (5) (0.66 g) was added to a solution of sodium carbonate (0.6 g) in water (120 ml) and dioxane (40 ml), then added dropwise at 0°C to FMOC-OSuc (0.87 g ) and dioxane (10 mL). After 2 hours at room temperature, the dioxane was removed in vacuo, and the mixture was acidified with hydrochloric acid. The solid was collected by filtration and dried under vacuum to give compound (6) (0.93 g, LCMS 442.1 [M+H]).

Preparation example 2 sees above-mentioned reaction scheme 1 step 1 : the preparation of resin 1

Argopore-MB-CHO resin (Argonaut Technologies, San Carlos, CA 94070) (10 g, 8.1 mmol) and EDC (45 mL) were combined and shaken for 5 minutes, followed by the addition of 4-fluoro-α-methylbenzylamine (5 gram). After shaking for 15 minutes, NaBH(OAc) ₃ (7.5 g) was added and shaking was continued for 20 hours at room temperature. The reaction mixture was transferred to a 250 mL flask, and methanol (50 mL) was added carefully. After the gas evolution stopped, the solvent was decanted, and the resin was washed successively with methanol (50 ml) containing 2N ammonium hydroxide, dichloromethane (100 ml), methanol (100 ml) and dichloromethane (2×100 ml) . The resin was collected by filtration and dried in a vacuum oven at 40°C. Step 2 : Preparation of Resin 2

Resin 1 (2 g) from Step 1 suspended in dichloromethane (10 mL) was added DIPEA (1.5 mL) and acid chloride (7) in dichloromethane (10 mL, 0.27 M, prepared from the corresponding acid (6 ) and oxalyl chloride in dichloromethane, prepared by reaction at room temperature). Shake at room temperature for 24 hours, then filter and wash with dichloromethane, methanol, THF, methanol, and dichloromethane, then dry overnight in a vacuum oven at 40°C. Step 3 : Preparation of Resin 3

Resin 2 from step 2 was treated with 20% piperidine in DMF for 20 minutes. The reaction mixture was filtered and the procedure repeated several times, then filtered and the resin was washed sequentially with THF, dichloromethane, methanol and dichloromethane, the resin was resuspended in dichloromethane, cyclohexanone (20 equiv) and NaBH(OAc) ₃ (5 equiv). Shaking at room temperature for 48 hours, followed by filtration and washing with methanol, dichloromethane, THF and dichloromethane afforded resin 3.

Example 1

General method:

The resin from Preparation 2, step 3 was treated with 10% TFA in dichloromethane for 31 hours. Filter and repeat this step. The organic layers are combined and concentrated to give the compound of formula I.

Using this method, the compounds shown in Table 1 were prepared from the appropriate ^R1 -CH( ^R3 ) ^-NHR4 amine and _R2 -containing aldehydes, ketones, alkyl halides or alkyl methanesulfonates. The compounds shown in Table 1 were identified by liquid chromatography-mass spectrometry using Sciex 100 equipment. The flow rate was 1 mL/min using a C18 column (3.3 cm x 4.6 mm, 3 microns in diameter, purchased from Supelco) with a gradient of 5% to 95% _CH3CN in water with 0.05% TFA over 10 minutes. The retention times (Rt) and the measured masses (equivalent to M+H) are listed in Table 1.

其中-CH(R¹)(R³)及R²的定义均如表1中所示：

The prepared compound has the following structural formula:

The definitions of -CH(R ¹ )(R ³ ) and R ² are as shown in Table 1:

步骤1：(R)-对-氟-α-甲基苄基醇Example 2

Step 1 : (R)-p-fluoro-α-methylbenzyl alcohol

Under nitrogen, borane dimethyl sulfide (14 mL, 0.14 mol) was added to a solution containing p-fluoroacetophenone (13.8 g, 0.1 mol) and (S)-3,3-diphenyl-1- In a cooled solution (0° C.) of methyltetrahydro-3H-pyrrolo-[1,2-c][1.3.2]oxazaborole (30 mL of a 2M solution in toluene) in anhydrous THF (400 mL). The solution was allowed to stir at 0°C for 30 minutes, then methanol (50 mL) was added, washed with 1M hydrochloric acid, water, saturated sodium bicarbonate, and dried over anhydrous sodium sulfate. Evaporation of the solvent gave 13.5 g of (R)-p-fluoro-α-methylbenzyl alcohol.

Analytical data: ¹ H NMR (CDCl ₃ ), δ 7.35 (d, 2H), δ 7.1 (t, 2H), δ 4.9 (q, 1H), δ 1.45 (d, 3H).

HPLC: (Chiralpak AS column, 10% isopropanol/hexane, flow rate 1 mL/min), 5.68 min R isomer 98%; 6.10 min S isomer 2%. Step 2 : (S)-p-fluoro-α-methylbenzyl azide

Diphenylphosphoryl azide (55.4 g, 0.395 mol) was added to a cooled solution (-15°C) of the product from Step 1 (23.5 g, 0.168 mol) in toluene (300 mL) followed by DBU (30.4 mL , 0.2 mol). After stirring overnight, the resulting biphasic solution was poured into 500 mL of 1N hydrochloric acid. The aqueous layer was extracted with toluene, and the combined organic layers were washed with water, 1N hydrochloric acid and dried over sodium sulfate. After removing the solvent, 25 g of crude product were obtained and used in the next step without purification. Step 3 : (S)-p-fluoro-α-methylbenzylamine

A mixture of the crude product from step 2 (1.2 g, 7.3 mmol), conc. HCl (1 mL in 50 mL methanol) and Pd/C (10% w/w) (140 mg) was hydrogenated at 60 PSi for 4 hours. After removing the solvent, the residue was redissolved in 20 mL of water and washed with ether. The aqueous layer was then basified to pH 11 and extracted with ether. The ether solution was dried and the solvent was evaporated to yield 1.1 g of (S)-p-fluoro-α-methylbenzylamine.

Analytical data: ¹ H NMR (CDCl ₃ ), δ 7.3 (dd 2H), δ 7.05 (t, 2H), δ 4.8 (q, 1H), δ 1.5 (d, 3H).

HPLC: (Chiralpak CR(+) column, perchloric acid in water, pH 1.5, flow rate 1 ml/min), 97.5% S isomer in 31.8 minutes, 2.5% R isomer in 43.2 minutes. Step 4 : N-[1-(S)-p-fluorophenethyl]-4-(4'-piperidinylmethyl)benzamide

EDCI hydrochloride (6.1 g, 34.4 mmol) was added to a solution containing the product of step 3 (4.8 g, 34.5 mmol), 4-(N-Boc-piperidinylmethyl)benzoic acid (10 g, 31.3 mmol mol) and DMAP (0.38 g, 3.1 mmol) in dichloromethane and the final solution was allowed to stir for 2 hours before quenching with 0.5N hydrochloric acid. The mixture was extracted with dichloromethane and washed with water. After removing the solvent, the residue was dissolved in TFA (10 mL) for 30 min. After removal of the acid, the residue was chromatographed to give 10.2 g of the desired product.

Analytical data: ¹ H NMR (CD3OD), δ7.76(d, 2H), δ7.40(dd, 2H), δ7.25(dd, 2H), δ7.05(t, 2H), δ5.2( q, 1H), δ2.95(m, 2H), δ2.59(m, 2H), δ2.5(m, 2H), δ1.65(m, 1H), δ1.6(m, 2H), δ1.55(d, 3H), δ1.95(m, 2H) step 5 :

Cyclohexanone (3.24 g, 33 mmol) and sodium triacetoxyborohydride (7.01 g, 33 mmol) were added to a solution of the product from step 4 (7.5 g, 22 mmol) in DCE (250 mL) middle. After the final mixture was stirred overnight, it was poured into 1N hydrochloric acid and the solution was washed with ether. The aqueous layer was basified to pH 11 with sodium hydroxide and extracted with dichloromethane. The organic layer was dried and the solvent was evaporated, and the residue was chromatographed to afford 8.5 g of the title compound.

Analytical data: ¹ H NMR (CD3OD): δ7.668 (d, 2H), δ7.354 (q, 2H),

δ7.186(d, 2H), δ7.028(t, 2H), δ5.309(m, 1H), δ2.858(d, 2H), δ2.565(d, 2H), δ2.231(m , 1H), δ2.105(td, 2H), δ1.86-1.75(m, 4H), δ1.614(m, 2H), δ1.58(d, 3H), δ1.56-1.42(m, 3H), δ1.32-1.02 (m, 6H).

[α] _D/20 in methanol 19.8.

C ¹³ (CDC ¹³ ) δ167.4, 164.2, 161.7, 145.9, 140.0, 132.9, 130.3, 128.9, 128.8, 127.8, 116.6, 116.4, 65.1, 50.4, 49.7, 44.4, 39.5, 33.9, 30.1, 27.6, 27 23.1

ES-LCMS, 6.36min, LC area 100%, M+1,423.

Melting point (hydrochloride): 234-235°C.

Example 3

The product (0.522 g, 1.537 mmol) and sodium carbonate (3.26 g, 30.74 mmol) in the example 2, step 4 were added to the solution containing t, t-3,5-dimethylcyclohexyl-p-toluenesulfonic acid The ester (1.35 g, 4.61 mmol) was dissolved in 4-methyl-2-pentanone (5 mL), and the mixture was refluxed overnight. The final reaction mixture was filtered and the solids were washed with dichloromethane. The combined organic layers were concentrated and the residue was chromatographed using a mixture of 5% v/v 2M ammonia in methanol and dichloromethane to afford 420 mg of the title compound.

Analytical data: ¹ H NMR (CDCl ₃ ): δ7.75(d, 2H), δ7.40(q, 2H), δ7.248(d, 2H), δ7.04 6(t, 2H), δ5. 217(q, 1H), δ2.918(d, 2H), 2.597(d, 2H), δ2.408(tt, 1H), δ2.241(t, 2H), δ1.835(d, 2H), δ1.7-1.57(m, 3H), δ1.542(d, 3H), δ1.5-1.22(m, 6H), δ0.92(d, 6H), δ0.835(q, 1H), δ0 .498(q, 1H).

Example 4 Step 1 :

Triphenylphosphine (0.33 g, 1.27 mmol) was added to a solution of c,c-3,5-bis-trifluoromethylcyclohexanol (0.2 g, 0.85 mmol) in anhydrous benzene (8 mL) , followed by the addition of DEAD (0.22 g, 1.27 mmol). The solution was stirred at room temperature for 5 minutes, then methyl tosylate (0.236 g, 1.27 mmol) was added. After the reaction was stirred at room temperature for 72 hours, it was diluted with ether (80 mL), washed with water, 1N hydrochloric acid, water and saturated sodium bicarbonate, brine, and dried over magnesium sulfate. The organic solvent was evaporated and the residue was chromatographed on a silica gel column with 10% ethyl acetate/hexanes to afford 0.28 g of the desired tosylate (85% yield). H ¹ NMR (CDCl ₃ ): δ7.8(d, 2H), δ7.4(d, 2H), δ5.0(b, 1H), δ2.5(m, 2H), δ2.45(s, 3H), δ2.18(m, 3H), δ1.45(m, 2H), δ1.35(m, 1H). Step 2 :

The same procedure of Example 3 was repeated, and using the tosylate of Step 1, the title compound was prepared.

H ¹ NMR (CDCl ₃ ): δ7.65(d, 2H), δ7.35(m, 2H), δ7.2(d, 2H), δ7.0(m, 2H), δ5.3(m, 1H), δ2.8(m, 2H), δ2.6(d, 2H), δ2.45(m, 1H), δ2.05-2.25(m, 6H), δ1.45-1.8(m, 4H ), δ 1.6 (d, 3H), δ 1.2-1.4 (m, 5H). ES-LCMS, Rt 6.11 min, measured mass 559 (M+H).

Example 5

Bicyclo[3.3.1]nonan-9-one (1.0 g, 7.2 mmol) and sodium triacetoxyborohydride (3.24 g, 14 mmol) were added to the product containing Example 2, Step 4 (2.46 g , 7.2 mmol) in a solution of DCE (75 ml). After the final mixture was stirred overnight, the solvent was evaporated and the residue was chromatographed to afford 0.640 g of the title compound.

Analytical data: ¹ H NMR (CD3OD): δ7.6 75 (d, 2H), δ7.360 (q, 2H),

δ7.194(d, 2H), δ7.027(t, 2H), δ5.309(m, 1H), δ3.07(d, 2H), δ2.56(d, 2H), δ2.00-1.66 (m, 11H), δ 1.58 (d, 3H), δ 1.54-1.43 (m, 7H), δ 1.33-1.23 (m, 4H).

Example 6

Sodium hydride (57.6 mg, 2.4 mmol) was suspended in DMF under nitrogen, followed by the addition of a DMF solution of the compound prepared in Example 2 (100 mg, 0.24 mmol) with stirring. After 30 min, _CH31 (0.017 mL, 0.264 mmol) was added and the mixture was stirred for 40 h. The mixture was poured into an ice-water bath, the precipitate was collected by filtration, the solid was washed with water and dried in vacuo to afford 100 mg of the title compound. LCMS: Rt 6.81 min, found mass 437 (M+H).

Example 7

13 (2 mmol) in dichloromethane (2 mL) was added to a solution of 12 (0.91 mmol) in a 1:1 mixture of water/saturated sodium bicarbonate (10 mL) with vigorous stirring. After 1 hour, the organic layer was separated, and the aqueous layer was washed three times with dichloromethane. The combined organic layers were washed with saturated sodium bicarbonate and brine, and dried over sodium sulfate. After removal of the solvent, the residue was chromatographed on a semi-preparative HPLC column with 2% (v/v) 7N _NH3 in methanol and dichloromethane to afford 140 mg of the desired product.

H ¹ NMR (CDCl ₃ ) δ8.55, d, 2H, δ7.7, d 2H, δ7.26d, 2H, δ7.20d, 2H; δ6.45d, 1H; δ5.3, m, 1H; δ2. 85, m 2H; δ2.58, d, 2H; δ2.25, m, 1H; δ2.1, m, 2H; δ1.7-1.9, m, 4H; .0-1.35, m; 6H.

LCMS: C18 reverse-phase column, 5%-95% gradient of CH ₃ CN/water within 10 minutes, retention time, 3.58 minutes, mass spectral data M+1 406.

Claims (10)

1. the compound of a following structural I

Or its pharmacy acceptable salt, ester or its solvate, wherein

R ¹Be R ⁵-(C ₃-C ₈) cycloalkyl, R ⁵-(C ₃-C ₈) cycloalkyl (C ₁-C ₆) alkyl, R ⁵-aryl, R ⁵-aryl-(C ₁-C ₆) alkyl or R ⁵-heteroaryl;

R ²Be H, (C ₁-C ₆) alkyl, R ⁶(C ₃-C ₈) cycloalkyl, R ⁶-(C ₃-C ₈) cycloalkyl-(C ₁-C ₆) alkyl, R ⁶-Heterocyclylalkyl, R ⁶-(C ₆-C ₁₀) bridge ring alkyl or R ⁶-bridge Heterocyclylalkyl;

R ³Be C ₁-C ₆Alkyl or-CH ₂OH;

R ⁴Be H or C ₁-C ₆Alkyl;

R ⁵Comprise H, C for independently being selected from ₁-C ₆Alkyl, halogen ,-OH, C ₁-C ₆Alkoxyl group ,-CF ₃,-CN ,-CO ₂R ⁴,-CONHR ⁴,-SO ₂NHR ⁴,-NHSO ₂R ⁴, and-NHC (O) R ⁴1-4 substituting group; And

R ⁶Comprise H, C for independently being selected from ₁-C ₆Alkyl, halogen ,-OH, C ₁-C ₆Alkoxyl group ,-CF ₃,-NH ₂, (C ₁-C ₆) alkylamino, phenyl, C ₁-C ₂Alkylenedioxy group, and (C ₁-C ₆) 1-4 substituting group of alkoxy carbonyl.

2. compound as claimed in claim 1, R wherein ¹Be R ⁵-phenyl or R ⁵-cyclohexyl.

3. compound as claimed in claim 2, R wherein ⁵Be H, halogen or C ₁-C ₆Alkyl.

4. compound as claimed in claim 1, R wherein ²Be R ⁶-C ₃-C ₈Cycloalkyl.

5. compound as claimed in claim 4, R wherein ⁶Be H or C ₁-C ₆Alkyl.

6. compound as claimed in claim 1, it is selected from following compounds

7. a pharmaceutical composition comprises that the compound as claimed in claim 1 for the treatment of significant quantity combines with pharmaceutically acceptable carrier.

8. compound as claimed in claim 1 is used to prepare the purposes of the medicine for the treatment of cognition or nervous system disease.

9. compound as claimed in claim 1 and acetyl cholinesterase enzyme and usefulness are used to prepare the purposes of the medicine of treatment cognition or nervous system disease.

10. medicine box that is used for the treatment of cognitive dissonance or nervous system disease; comprise that the single-section charge compounds that will be used in combination is placed in the container separately; be the compound of claim 1 in the one container; it in second container acetyl cholinesterase enzyme inhibitors; all in pharmaceutically acceptable carrier, its binding capacity is a significant quantity for they.